The present invention relates to ligand involved in B lymphocyte activation and proliferation, TALL-1, and to its receptor, TALL-1 receptor. More particularly, the present invention relates to TALL-1 and TALL-1 receptor, to homologues of such proteins, to nucleic acid molecules encoding such proteins, and to methods of making and using such proteins.
Members of the tumor necrosis factor (TNF) ligand family play important roles in various physiological and pathological processes, including cell proliferation, differentiation, apoptosis, modulation of immune response and induction of inflammation (Smith et al., (1994), Cell 76:959-962; Grewal et al., (1996), The role of CD40 ligand in immunity and auto-immunity. Page 125-134. In Immune Tolerance, editors: J. Banchereau, B. Dodet, R.
Schwartz, E. Trannoy. Elsevier Press: Paris.; Nataga, (1997), Cell 88:355-365; Baker et al., (1998), Onocogene 17:3261-3270; Ashkenazi et al., (1999), Curr. Opin. Cell Biol. 11:255-260). At least sixteen members of the TNF ligand family have been identified. These include TNF, FasL, Lymphotoxin-xcex1, Lymphotoxin-xcex2, TRA1L/APO-2L, CD27L, CD30L, CD40L, 4-1BBL, OX40L, TRANCE/RANKL, LIGHT, TWEAK, TL 1, APRIL/TALL-2 and TALL-1 (Smith et al., (1994), Cell 76:959-962; Grewal et al., (1996), The role of CD40 ligand in immunity and auto-immunity. Page 125-134. In Immune Tolerance, editors: J. Banchereau, B. Dodet, R. Schwartz, E. Trannoy. Elsevier Press: Paris.; Nataga, (1997), Cell 88:355-365; Baker et al., (1998), Onocogene 17:3261-3270; Ashkenazi et al., (1999), Curr. Opin. Cell Biol. 11:255-260; Shu et al., (1999), J. Leukocyte Biology65:680-683; Schneider et al., (1999), J. Exp. Med. 189:1747-1756; Moore et al., (1999), Science 285:260-263; Mukhopadhyay et al., (1 999), J. Biol. Chem. 274:15978-15981). Most TNF family members are synthesized as type II transmembrane precursors. Their extracellular domains can be cleaved by metalloproteinases to form soluble cytokines. The soluble and membrane-bound TNF ligand family members bind to receptors belonging to the TNF receptor family, which are type I transmembrane proteins with characteristic cysteine rich motifs (Smith et al., (1994), Cell 76:959-962; Nataga, (1997), Cell 88:355-365; Baker et al., (1998), Oncogene 17:3261-3270; Ashkenazi et al., (1999), Curr. Opin. Cell Biol. 11:255-260).
Members of the TNF family interact with their cognate receptors either through cell-cell interactions or as soluble proteins after their extracellular domains are cleaved by metalloproteinases (Smith et al., (1 994),Cell 76:959-962; Nataga, (1 997), Cell 88:355 -365). The TNF receptor family contains about 20 members. Stimulation of TNF receptor family members by their ligands triggers overlapping and divergent intracellular signal transduction pathways, including those that lead to apoptosis, NF-xcexaB and AP1 activation (Smith et al., (1994), Cell 76:959-962; Nataga, (1997), Cell 88:355-365). Therefore regulation of TNF family members that specifically affect immune responses is desirable.
A wide variety of medical treatments require regulation of the immune response in a patient. Such treatments include, for example, vaccinations, treatments for autoimmune diseases, immunodeficiency diseases, immunoproliferative diseases, and treatments involving the transplantation of organs and skin. Traditional reagents and methods used to regulate a subject""s immune response often results in unwanted side effects. For example, immunosuppressive reagents such as cyclosporin A, azathioprine, and prednisone are used to suppress the immune system of a patient with an autoimmune disease or patients receiving transplants. Such reagents, however, suppress a patient""s entire immune response, thereby crippling the ability of the patient to mount an immune response against infectious agents not involved in the original disease. Due to such harmful side effects and the medical importance of immune regulation, reagents and methods to regulate specific parts of the immune system have been the subject of study for many years.
The present invention can be used to overcome traditional problems with immunoregulatory reagents by more specifically regulating cells, ligands and receptors of the immune system in vivo.
The present invention generally relates to TALL-1 nucleic acid molecules, proteins (including homologues), antibodies, and methods of making and using the same. The present invention also relates to TALL-1 receptor nucleic acid molecules, proteins (including homologues), antibodies, and methods of using the same. In particular, the present invention relates to methods of regulating the interaction between TALL-1 and the TALL-1 receptor to regulate monocyte, macrophage and B lymphocyte mediated immune responses.
One embodiment of the present invention relates to an isolated TALL-1 protein. Preferably, the isolated TALL-1 protein comprises an amino acid sequence selected from the group of: (a) SEQ ID NO:2; and (b) a homologue of SEQ ID NO:2. The homologue comprises an amino acid sequence selected from the group of: (i) an amino acid sequence that is at least about 40% identical to SEQ ID NO:2 over at least 175 amino acids of SEQ ID NO:2; and, (ii) an amino acid sequence that is at least about 40% identical to SEQ ID NO:2 over positions 134-285 of SEQ ID NO:2. In a more preferred embodiment, the protein comprises an amino acid sequence selected from the group of: (a) an amino acid sequence that is at least about 60% identical to SEQ ID NO:2 over at least 175 amino acids of SEQ ID NO:2; and, (b) an amino acid sequence that is at least about 60% identical to SEQ ID NO:2 over positions 134-285 of SEQ ID NO:2. In an even more preferred embodiment, the protein comprises an amino acid sequence selected from the group of: (a) an amino acid sequence that is at least about 80% identical to SEQ ID NO:2 over at least 175 amino acids of SEQ ID NO:2; and, (b) an amino acid sequence that is at least about 80% identical to SEQ ID NO:2 over positions 134-285 of SEQ ID NO:2. In an even more preferred embodiment, the protein comprises an amino acid sequence selected from the group consisting of: (a) an amino acid sequence that is at least about 90% identical to SEQ ID NO:2 over at least 175 amino acids of SEQ ID NO:2; and, (b) an amino acid sequence that is at least about 90% identical to SEQ ID NO:2 over positions 134-285 of SEQ ID NO:2.
In another embodiment, the isolated TALL-1 protein of the present invention has an amino acid sequence comprising at least about 30 contiguous amino acids of SEQ ID NO:2. Preferably, the protein has an amino acid sequence comprising at least about 50 contiguous amino acids of SEQ ID NO:2, and more preferably, at least about 130 contiguous amino acids of SEQ ID NO:2. In a particularly preferred embodiment, the protein comprises amino acid residues 134 through 285 of SEQ ID NO:2. In another particularly preferred embodiment, the protein comprises an amino acid sequence represented by SEQ ID NO:2.
In another embodiment, the isolated TALL-1 protein of the present invention is encoded by a nucleic acid molecule comprising a nucleic acid sequence that hybridizes under low stringency conditions, and preferably, moderate stringency conditions, and more preferably, high stringency conditions, to the complement of a nucleic acid sequence represented by SEQ ID NO:1. In a particularly preferred embodiment, the protein is encoded by a nucleic acid molecule comprising a nucleic acid sequence represented by SEQ ID NO:1. In another particularly preferred embodiment, the protein is encoded by a nucleic acid molecule comprising a nucleic acid sequence represented by nucleotide positions 402 through 855 of SEQ ID NO:1.
In one embodiment of the present invention, an isolated TALL-1 protein is soluble. In another embodiment, the isolated TALL-1 protein is a membrane protein. Preferably, the isolated TALL-1 protein of the present invention has TALL-1 biological activity. Such biological activities include, but are not limited to: (a) binding to a TALL-1 receptor; (b) costimulation of B lymphocyte proliferation; (c) costimulation of B lymphocyte activation; and/or (d) support of B lymphocyte survival. Preferably, the isolated TALL-1 protein binds to a TALL-1 receptor. In one embodiment, a TALL-1 protein binds to an activates a TALL-1 receptor. In another embodiment, the TALL-1 protein is a TALL-1 homologue that does not activate a TALL-1 receptor.
Yet another embodiment of the present invention relates to an isolated antibody that specifically binds to the isolated TALL-1 protein of the present invention.
Another embodiment of the present invention relates to an isolated nucleic acid molecule comprising a nucleic acid sequence encoding a protein comprising an amino acid sequence selected from the group of: (a) SEQ ID NO:2; and, a homologue of SEQ ID NO:2. The homologue comprises an amino acid sequence selected from the group consisting of: (i) an amino acid sequence comprising at least 130 contiguous amino acids of SEQ ID NO:2; and, (ii) an amino acid sequence comprising at least positions 134-285 of SEQ ID NO:2. Preferably, the homologue comprises an amino acid sequence comprising at least 150 contiguous amino acids of SEQ ID NO:2, and even more preferably, the homologue comprises an amino acid sequence comprising at least 175 contiguous amino acids of SEQ ID NO:2. In a particularly preferred embodiment, the nucleic acid sequence encodes a protein comprising an amino acid sequence represented by SEQ ID NO:2, or a protein comprising an amino acid sequence comprising amino acids 134-285 of SEQ ID NO:2.
Preferably, the isolated nucleic acid sequence comprises at least 390 contiguous nucleotides of SEQ ID NO:1. In another aspect, the nucleic acid sequence hybridizes under low stringency conditions, and preferably, moderate stringency conditions, and more preferably, high stringency conditions, to the complement of a nucleic acid sequence SEQ ID NO:1. In a particularly preferred embodiment, the nucleic acid sequence comprises SEQ ID NO:1, or the nucleic acid sequence comprises nucleotides 402 through 855 of SEQ ID NO:1.
One embodiment of the present invention relates to an isolated nucleic acid molecule as set forth above, operatively linked to a transcription control sequence. Another embodiment of the present invention relates to a recombinant cell comprising an isolated nucleic acid molecule as set forth above, wherein the cell expresses the recombinant nucleic acid molecule. Yet another embodiment of the present invention relates to a recombinant virus, comprising an isolated nucleic acid molecule as set forth above.
Yet another embodiment of the present invention relates to a recombinant nucleic acid molecule comprising an expression vector operatively linked to a nucleic acid molecule comprising a nucleic acid sequence encoding a protein having TALL-1 biological activity. The protein comprises an amino acid sequence selected from the group of: (a) SEQ ID NO:2; and, (b) a homologue of SEQ ID NO:2, wherein the homologue comprises an amino acid sequence selected from the group of: (i) an amino acid sequence that is at least about 40% identical to SEQ ID NO:2 over at least 175 amino acids of SEQ ID NO:2; and, (ii) an amino acid sequence that is at least about 40% identical to SEQ ID NO:2 over positions 134-285 of SEQ ID NO:2.
Another embodiment of the present invention relates to a method to produce a TALL-1 protein. Such a method includes the step of culturing an isolated cell to express any of the recombinant nucleic acid molecules described above, under conditions whereby a TALL-1 protein encoded by the recombinant nucleic acid molecule is produced.
Yet another embodiment of the present invention relates to an isolated TALL-1 receptor homologue, wherein the homologue comprises an amino acid sequence that is: (a) at least about 40% identical to SEQ ID NO:11 over at least 35 amino acids of SEQ ID NO:11; and, (b) less than 100% identical to an amino acid sequence selected from the group of SEQ ID NO:11 and SEQ ID NO:17. In a preferred embodiment, the TALL-1 receptor homologue is at least about 65% identical to SEQ ID NO:1, and more preferably, the homologue is at least about 75% identical to SEQ ID NO:11, and more preferably, the homologue is at least about 90% identical to SEQ ID NO:11. In one embodiment of the present invention, the homologue is less than about 95% identical to the amino acid sequence selected from the group of SEQ ID NO:11 and SEQ ID NO:17; in another embodiment, the homologue is less than about 90% identical to the amino acid sequence selected from the group consisting of SEQ ID NO:11 and SEQ ID NO:17.
In one embodiment, the TALL-1 receptor homologue encoded by a nucleic acid molecule comprising a nucleic acid sequence that hybridizes under moderate stringency conditions to the complement of SEQ ID NO:10, and more preferably, the homologue is encoded by a nucleic acid molecule comprising a nucleic acid sequence that hybridizes under high stringency conditions to the complement of SEQ ID NO:10.
In one embodiment, the TALL-1 receptor homologue binds to TALL-1. In one embodiment, the homologue is a soluble TALL-1 receptor. In another embodiment, the homologue does not bind to TALL-1. In another embodiment, the homologue activates NF-xcexaB in a cell expressing the homologue through a TRAF5, TRAF6, NIK, IKKxcex1 and IKKxcex2 dependent pathway. In one embodiment, the homologue costimulates B lymphocyte proliferation in a B lymphocyte expressing the homologue.
Yet another embodiment of the present invention relates to an isolated nucleic acid molecule comprising a nucleic acid sequence encoding a TALL-1 receptor homologue. The homologue comprises an amino acid sequence that is: (a) at least about 40% identical to SEQ ID NO:11 over at least 35 amino acids of SEQ ID NO:11; and, (b) less than 100% identical to an amino acid sequence selected from the group of SEQ ID NO:11 and SEQ ID NO:17.
Another embodiment of the present invention relates to a composition comprising: (a) a compound; and (b) a pharmaceutically acceptable carrier. The compound of (a) is selected from the group of: (i) an isolated TALL-1 protein; (ii) an isolated antibody that selectively binds to the TALL-1 protein of (i); (iii) an isolated TALL-1 receptor; and, (iv) an isolated antibody that selectively binds to the TALL-1 receptor of (iii).
In one aspect of the composition of the present invention, the compound is an isolated TALL-1 protein. Such a TALL-1 protein can be any heretofore described TALL-1 protein, including a protein comprising an amino acid sequence selected from the group of: (a) SEQ ID NO:2; and, (b) a homologue of SEQ ID NO:2, wherein the homologue comprises an amino acid sequence selected from the group of: (i) an amino acid sequence that is at least about 40% identical to SEQ ID NO:2 over at least 175 amino acids of SEQ ID NO:2; and, (ii) an amino acid sequence that is at least about 40% identical to SEQ ID NO:20 over positions 134-285 of SEQ ID NO:2. In one embodiment, the isolated TALL-1 protein is encoded by a nucleic acid sequence that hybridizes under moderate stringency conditions to the complement of a nucleic acid sequence encoding SEQ ID NO:2. In another embodiment, the isolated TALL-1 protein comprises an amino acid sequence represented by SEQ ID NO:2.
In another aspect of he composition of the present invention, the compound is an isolated TALL-1 receptor. In one embodiment, the receptor is a soluble TALL-1 receptor. In another embodiment, the receptor comprises an amino acid sequence selected from the group of: (a) SEQ ID NO:1; and, (b) a TALL-1 receptor homologue comprising an amino acid sequence that is: (i) at least about 40% identical to SEQ ID NO:11 over at least 35 amino acids of SEQ ID NO:11; and, (ii) less than 100% identical to an amino acid sequence selected from the group of SEQ ID NO:11 and SEQ ID NO:17.
Yet another embodiment of the present invention relates to a method to activate a TALL-1 receptor, comprising contacting a TALL-1 receptor with a TALL-1 protein. Preferably, the TALL-1 protein comprises an amino acid sequence selected from the group of: (a) SEQ ID NO:2; and, (b) a homologue of SEQ ID NO:2, wherein the homologue comprises an amino acid sequence selected from the group of: (i) an amino acid sequence that is at least about 40% identical to SEQ ID NO:2 over at least 175 amino acids of SEQ ID NO:2; and, (ii) an amino acid sequence that is at least about 40% identical to SEQ ID NO:2 over positions 134-285 of SEQ ID NO:2.
Another embodiment of the present invention relates to a method to regulate B lymphocyte proliferation, activation, and/or survival, comprising regulating the biological activity of a TALL-1 receptor expressed by a B lymphocyte. In one aspect, the step of regulating comprises inhibiting the biological activity of the TALL-1 receptor. For example, the step of regulating can include inhibiting the interaction of TALL-1 with the TALL-1 receptor. In one aspect of this embodiment, the step of inhibiting comprises contacting TALL-1 expressed by a monocyte or macrophage with a compound that inhibits binding of the TALL-1 to the TALL-1 receptor. Such a compound can include, but is not limited to: (a) an antibody that selectively binds to TALL-1 and inhibits the binding of TALL-1 to the TALL-1 receptor; (b) a soluble TALL-1 receptor; and, (c) a TALL-1 receptor homologue, wherein the homologue comprises an amino acid sequence that is at least about 40% identical over at least 35 amino acids to SEQ ID NO:11, wherein the homologue binds to TALL-1, and wherein the receptor does not activate NF-xcexaB when expressed by a cell. In one aspect, the compound is preferably a soluble TALL-1 receptor.
In another aspect of this embodiment, the step of inhibiting comprises contacting the TALL-1 receptor with a compound that inhibits the interaction of TALL-1 with the TALL-1 receptor. Such a compound can include, but is not limited to: (a) an antibody that selectively binds to the TALL-1 receptor and inhibits the binding of the TALL-1 receptor by TALL-1 ; and/or (b) a TALL-1 homologue, wherein the homologue comprises an amino acid sequence selected from the group of: (i) an amino acid sequence that is at least about 40% identical to SEQ ID NO:2 over at least 175 amino acids of SEQ ID NO:2; and, (ii) an amino acid sequence that is at least about40% identical to SEQ ID NO:2 over positions 134-285 of SEQ ID NO:2. In this aspect, the homologue binds to the TALL-1 receptor but does not activate the TALL-1 receptor.
In another aspect of the method of the present invention, the step of regulating comprises stimulating the biological activity of the TALL-1 receptor. In one aspect, the method includes the step of contacting the TALL-1 receptor with a compound that increases TALL-1 receptor activity. Such a compound can include, but is not limited to: (a) an isolated TALL-1 protein; and, (b) an antibody that selectively binds to the TALL-1 receptor and activates the receptor. Preferably, the isolated TALL-1 protein comprises an amino acid sequence selected from the group of: (a) SEQ ID NO:2; and, (b) a homologue of SEQ ID NO:2, wherein the homologue comprises an amino acid sequence selected from the group of: (i) an amino acid sequence that is at least about 40% identical to SEQ ID NO:2 over at least 175 amino acids of SEQ ID NO:2; and, (ii) an amino acid sequence that is at least about 40% identical to SEQ ID NO:2 over positions 134-285 of SEQ ID NO:2.
In one aspect of the method of the present invention, the regulation of B lymphocyte proliferation, activation and/or survival by the method is effective to regulate a B-lymphocyte immune response in an animal. In another aspect of the method of the present invention, the inhibition of B lymphocyte proliferation, activation and/or survival by the method is effective to inhibit a B lymphocyte-associated autoimmune disease.
Yet another embodiment of the present invention relates to a method to inhibit a B lymphocyte immune response in a patient, comprising inhibiting the interaction of TALL-1 with a TALL-1 receptor in the patient. Another embodiment of the present invention relates to a method to inhibit a B lymphocyte-associated autoimmune disease in a patient, comprising inhibiting the interaction of TALL-1 with a TALL-1 receptor in the patient.
Yet another embodiment of the present invention relates to a method to identify compounds that regulate the interaction between TALL-1 and a TALL-1 receptor. Such a method includes the steps of: (a) contacting a TALL-1 receptor with a putative regulatory compound; (b) contacting the TALL-1 receptor with a TALL-1 protein comprising an amino acid sequence selected from the group of: (i) SEQ ID NO:2; and, (ii) a homologue of SEQ ID NO:2, wherein the homologue comprises an amino acid sequence selected from the group of: (1) an amino acid sequence that is at least about 40% identical to SEQ ID NO:2 over at least 175 amino acids of SEQ ID NO:2; and, (2) an amino acid sequence that is at least about 40% identical to SEQ ID NO:2 over positions 134-285 of SEQ ID NO:2 or a homologue of TALL-1; and (c) detecting whether the putative regulatory compound regulates the TALL-1 receptor. The TALL-1 protein binds to and activates the TALL-1 receptor in the absence of the putative regulatory compound. In one aspect, the step of detecting comprises detecting whether the putative regulatory compound inhibits the binding of the TALL-1 protein to the TALL-1 receptor. In another aspect, the step of detecting comprises detecting whether the putative regulatory compound inhibits the activation of the TALL-1 receptor as compared to in the absence of the putative regulatory compound.